Volume 11, Issue 2, November – December 2011; Article-028
ISSN 0976 – 044X
Research Article
BIOCHEMICAL EVALUATION OF ANTIDIABETIC, ANTILIPIDEMIC AND ANTIOXIDANT NATURE OF
CASSIA AURICULATA SEEDS STUDIED IN ALLOXAN-INDUCED EXPERIMENTAL DIABETES IN RATS
*
Subramanian S , Uma S K and Sriram Prasath G
Department of Biochemistry, University of Madras, Guindy Campus, Chennai 600 025, India.
*Corresponding author’s E-mail: subbus2020@yahoo.co.in
Accepted on: 16-09-2011; Finalized on: 30-11-2011.
ABSTRACT
Cassia auriculata Linn. a member of genus Cassia belongs to family Caesalpiniaceae is commonly known as Tanner’s cassia. Various
parts of the plant have been reported to possess a number of therapeutic activities to manage several diseases. The flowers and
seeds are reported to be useful in the treatment of diabetes. Hence, the present study was designed to evaluate the hypoglycemic,
hypolipidemic and antioxidant properties of Cassia auriculata seed extract in alloxan-induced experimental diabetic rats. The effects
of oral administration of Cassia auriculata seed extract (400 mg/kg b.w.) on the levels of biochemical parameters were determined
in experimental groups of rats. The altered levels of biochemical parameters in the diabetic rats were significantly reverted back to
near basal values by the administration of Cassia auriculata seed extract for 30 days. The levels of lipid peroxides in the plasma and
pancreatic tissues of diabetic rats were elevated significantly and were normalized by the administration of Cassia auriculata seed
extract. The activities of pancreatic enzymic antioxidants and the levels of plasma non-enzymic antioxidants were markedly declined
in the diabetic rats. Upon treatment with Cassia auriculata seed extract to diabetic rats, these decreased levels were elevated to
near normal values. The altered levels of lipid profile were reverted back to near normalcy upon the extract treatment. The results
of the study indicate that Cassia auriculata seed extract possesses antilipidemic, antioxidant effects in addition to antidiabetic
activity. The results are comparable with glyclazide, an oral standard hypoglycemic drug. The phytochemicals present in the Cassia
auriculata seed extract may account for the observed pharmacological properties.
Keywords: Cassia auriculata, antidiabetic, antilipidemic, antioxidant nature.
INTRODUCTION
Diabetes mellitus is a chronic metabolic disorder resulting
from insulin deficiency and action. It is characterized by
hyperglycemia, altered metabolism of carbohydrates,
proteins and lipids1. The disease is associated with the
development of secondary complications as neuropathy,
renal failure, vision loss, macrovascular diseases and
amputations2. Treatment with oral hypoglycemic agents
is often associated with side effects related to
pharmacokinetic properties, secondary failure rates,
hypoglycemia, gastrointestinal disturbances, skin
reactions, hematological disorders, and rise in hepatic
enzyme levels3. Hence, the focus has been shifted to treat
various ailments through plant-derived drugs due to their
safety, efficacy, cultural acceptability and lesser side
effects.
The medicinal value of plants lies in some chemical
substances that produce a definite physiological action on
the human body. The most important of these bioactive
compounds of plants are alkaloids, flavonoids, tannins
4
and phenolic compounds . The WHO considers
phytotherapy in its health programs suggested basic
procedures for validation of drugs from plants origin in
developing countries.
Cassia auriculata Linn a member of genus Cassia
belonging to family caesalpiniaceae is commonly known
5
as Tanner’s cassia . The plant is also known as Avaram in
Tamil language or Avartaki in vernacular. The various
parts of plant have been reported to possess a number of
therapeutic activities to manage disease states like
leprosy, asthma, gout, rheumatism6 and diabetes7. It is
also used as antipyretic, antiulcer and in the treatment of
skin infections8. The plant has been reported to possess
hepatoprotective9, antiperoxidative and antihyper
glycemic activity10 and microbicidal activity11.
In the absence of systemic studies in the literature, the
present study was aimed to evaluate the hypoglycemic,
hypolipidemic and antioxidant properties of Cassia
auriculata seeds in alloxan induced experimental diabetes
in rats.
MATERIALS AND METHODS
Plant material
The seeds of Cassia auriculata were procured and
authenticated by a qualified taxonomist. A voucher
specimen was deposited at the herbarium, University of
Madras, Chennai.
Preparation of Plant extract
The Cassia auriculata seeds were dried at room
temperature and powdered in an electrical grinder, which
is then stored in an airtight container at 5°C until further
use. The seed powder was delipidated with Petroleum
Ether (60 - 80°C) for overnight. It was filtered and
Soxhalation extracted the residue with 95% Ethanol.
Ethanol was evaporated in a rotary evaporator at 40 50°C under reduced pressure.
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Volume 11, Issue 2, November – December 2011; Article-028
Preliminary phytochemical screening
The ethanolic extract of Cassia auriculata seeds were
subjected to preliminary phytochemical screening of
12,13
various plant constituents .
Experimental animals
Male albino wistar rats (150-180 g) were purchased from
TANUVAS, Madavaram, Chennai. The rats were housed in
polypropylene cages lined with husk and kept in Animal
house, Department of Biochemistry. It was renewed
every alternate days. The rats were fed with commercial
pellated rats chow (Lipton Ltd., Bangalore, India) and had
free access to water. The experimental rats were
maintained in a controlled environment (12:12 hours
light/dark cycle) and temperature (30 ± 2°C). The
experiments were designed and conducted in accordance
with the ethical norms approved by Ministry of Social
Justices and Empowerment, Government of India and
Institutional Animal Ethics Committee Guidelines for the
investigation of experimental pain in conscious rats. The
rats were acclimatized for one week before starting the
experiments.
Experimental design
The rats were grouped into 4 groups, comprising of 6 rats
in each group as follows:
Group I : Control rats (Water and food ad libitum)
Group II : Alloxan induced diabetic Rats
Group III: Diabetic rats treated with Cassia auriculata
seed extract (400 mg/Kg Body weight/day) in aqueous
solution orally for 30 days.
Group IV: Diabetic rats treated with glyclazide
(5mg/KgBody weight/day) in aqueous solution orally for
30 days.
During the experimental period, body weight and blood
glucose levels of all the rats were determined at regular
intervals. At the end of the experimental period, the rats
were fasted over night, anaesthetized, and sacrificed by
cervical decapitation. The blood was collected with and
without anticoagulant for plasma and serum separation
respectively.
Preparation of tissue homogenate
The pancreatic tissues were excised, rinsed in ice- cold
saline. Known amount of the tissues were homogenized
in Tris–HCl buffer (100 mM, pH 7.4) at 4°C, in a Potter–
Elvehjem homogenizer with a Teflon pestle at 600 rpm for
3 min. The homogenate was then centrifuged at 12,000×g for 30 min at 4°C. The supernatant was collected as
tissue homogenate, which was used to assay various
parameters.
Blood glucose level was estimated by the method of
glucose oxidase/peroxidase as described by Trinde14 and
urea by Natelson et al.15 Plasma was separated and used
for insulin assay using ELISA kit for rats. Levels of
hemoglobin and glycosylated hemoglobin were estimated
ISSN 0976 – 044X
16
according to methods of Drabkin and Austin and Nayak
and Pattabiraman17, respectively. Plasma was used for
protein assay18 and serum for determination of
19
20
creatinine and uric acid .
The pancreatic tissue homogenate was then centrifuged
at 5000g to remove cellular debris and supernatant was
used for the determination of lipid peroxides and
enzymatic antioxidants. Lipid peroxides were determined
using thiobarbituric acid reactive substances by the
21
method of Ohkawa et al. Levels of vitamin C, vitamin E,
ceruloplasmin and glutathione (GSH) were determined by
the methods of Omaye et al,22 Desai23, Ravin24, Sedlak and
25
Lindsay , respectively. Enzymatic antioxidants such as
superoxide
dismutase26,
catalase27,
glutathione
28
peroxidase in pancreatic supernatant.
Oral Glucose Tolerance Test (OGTT)
At the end of the experimental period, a fasting blood
sample was collected from all the groups of rats to
perform oral glucose tolerance test, rats were fasted for
12 h before the test and 2 g/kg glucose solution was
administered orally. Blood samples were taken by
severing the tip of the tail 1 h before and at 30, 60, 90 and
120 minutes after glucose administration. Blood glucose
was determined using ortho toluidine reagent.
Lipid profile
Plasma was used for the estimation of lipid profile.
Cholesterol content was estimated by the method of
Parekh and Jung29. Triglyceride was estimated by the
method of Rice30. HDL Cholesterol fraction was separated
by the precipitation techniques of Burstein and
Scholnick31 and the cholesterol content was determined
by method of Parekh and Jung29.
RESULTS
The preliminary results of the study revealed that Cassia
auriculata seed extract at the dose of 400 mg/Kg body
weight significantly normalize the elevated blood glucose
level, body weight and restored biochemical parameters
towards near normalcy.
Table 1 shows the qualitative analysis of phytochemical in
the ethanolic extract of Cassia auriculata seeds.
Phytochemical evaluation indicated the presence of
alkaloids, flavonoids, proteins, carbohydrates, saponins,
tannins, glycosides and phenols.
Fig 1 shows the changes in the levels of blood glucose,
after oral administration of glucose (3g/Kg) in normal
control and experimental group of rats. The data of OGTT
revealed that the blood glucose value in normal control
rat reach peak at 60 minutes after the oral glucose load
and gradually reverted back to near normal levels after
120 minutes. In diabetic control rats, the peak increase in
blood glucose concentration was observed after 60
minutes and remained high over the next 60 minutes.
Treated group showed definite lower peak blood glucose
values, 60 minutes after glucose load also gives lower
values almost at the end of 120 minutes.
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Volume 11, Issue 2, November – December 2011; Article-028
Table 1: Phytochemical screening of the ethanolic extract
of Cassia auriculata seeds
Phytoconstituents
Inference
Alkaloids
+
Flavonoids
+
Carbohydrates
+
Glycosides
Saponins
Tannins
+
Phytosterol
+
Triterpenoids
Proteins
Aminoacids
+
Anthraquinones
+
Phenols
+
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Figure 1: Effect of Cassia auriculata seed extract on (GTT)
in normal and alloxan induced diabetic rats
Values are given as mean ± SD for groups of six rats in each. Values are
statistically significant at p < 0.05. Statistical significance was compared
@
within the groups as follows: *compared with control, compared with
diabetic rats.
Table 2: Effect of C.auriculata extract on the levels of biochemical parameters in the experimental groups of rats
Groups
Glucose (mg/dl)
Insulin (µU/ml)
Hemoglobin (g/dl)
Glycosylated
hemoglobin (%)
Urine sugar
Control
98.79 ± 11.35
15.88 ± 2.85
15.85 ± 1.46
6.67 ± 1.52
Nil
Diabetic
Diabetic + C.auriculata extract
Diabetic + gliclazide
297.78 ± 21.46*
@
147.22 ± 13.54
@
126.12 ± 15.57
6.74 ± 1.12*
@
11.51 ± 1.45
@
12.54 ± 1.78
11.18 ± 1.67*
@
13.45 ± 2.39
@
13.90 ± 2.04
14.68 ± 2.39*
@
7.82 ± 1.94
@
7.59 ± 2.03
+++
Nil
Nil
Values are given as mean ± SD for groups of six rats in each. Values are statistically significant at p < 0.05. Statistical significance was compared within
@
the groups as follows: *compared with control, compared with diabetic rats. +++ indicates more than 2% sugar.
Table 3: Effect of C. auriculata seed extract on the levels of protein, urea, creatinine and uric acid in experimental groups of rats
Groups
Control
Diabetic
Protein (g/dl)
8.02 ± 1.05
5.36 ± 0.72*
Diabetic + C.auriculata extract
Diabetic + gliclazide
6.75 ± 0.78
@
7.22 ± 0.85
Urea (mg/dl)
23.91 ± 3.05
46.62 ± 3.98*
@
Creatinine (mg/dl)
1.06 ± 0.10
2.22 ± 0.22*
@
@
33.19 ± 3.42
@
31.82 ± 3.08
1.31 ± 0.10
@
1.30 ± 0.09
Uric acid (mg/dl)
2.57 ± 0.82
5.16 ± 1.38*
@
3.24 ± 0.86
@
2.76 ± 1.04
Values are given as mean ± SD for groups of six rats in each. Values are statistically significant at p < 0.05. Statistical significance was compared within
@
the groups as follows: *compared with control, compared with diabetic rats.
Table 4: Effect of C.auriculata extract on the level of TBARS in plasma and pancreas experimental groups of rats
TBARS
Groups
Plasma
Pancreas
Control
4.27 ± 0.95
40.14 ± 3.95
Diabetic
8.02 ± 1.51*
79.26 ± 8.44*
@
@
Diabetic + C.auriculata extract
5.39 ± 1.06
59.29 ± 6.71
@
@
Diabetic + gliclazide
5.14 ± 1.01
54.99 ± 7.33
Units: mM/100 g in tissues; nM/ml in plasma. Values are given as mean ± SD for groups of six rats in each. Values are statistically significant at p < 0.05.
@
Statistical significance was compared within the groups as follows: *compared with control, compared with diabetic rats.
Table 5: Effect of C. auriculata seed extract on the levels of non enzymatic antioxidants in plasma of experimental groups of rats
Groups
Vitamin C
Vitamin E
Ceruloplasmin
GSH
Control
1.50 ± 0.16
0.68 ± 0.08
12.30 ± 1.52
31.84 ± 3.99
Diabetic
0.51 ± 0.08*
0.33 ± 0.03*
5.03 ± 0.90*
15.99 ± 2.25*
Diabetic + C.auriculata extract
Diabetic + gliclazide
@
0.97 ± 0.13
@
1.01 ± 0.13
@
0.56 ± 0.08
@
0.60 ± 0.06
@
9.85 ± 1.36
@
10.41 ± 1.78
@
21.86 ± 2.97
@
24.15 ± 3.06
Units: mg/dl. Values are given as mean ± SD for groups of six rats in each. Values are statistically significant at p < 0.05. Statistical significance was
@
compared within the groups as follows: *compared with control, compared with diabetic rats.
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Volume 11, Issue 2, November – December 2011; Article-028
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Table 6: Effect of C.auriculata seed extract on the activity of SOD, Catalase and GPx, in pancreas of experimental groups of rats
Groups
Control
Diabetic
Diabetic + C.auriculata extract
Diabetic + gliclazide
SOD
5.36 ± 1.25
1.49 ± 0.31*
@
3.71 ± 0.82
@
3.98 ± 0.87
Catalase
15.82 ± 2.19
6.13 ± 1.40*
@
12.85 ± 1.99
@
13.06 ± 1.95
GPx
6.26 ± 1.01
3.09 ± 0.31*
@
4.96 ± 0.63
@
5.39 ± 0.89
Activity is expressed as: 50% of inhibition of epinephrine autoxidation/min for SOD; µM of hydrogen peroxide decomposed/min/mg of protein for
catalase; µM of glutathione oxidized/min/mg of protein for GPx. Values are given as mean ± SD for groups of six rats in each. Values are statistically
@
significant at p < 0.05. Statistical significance was compared within the groups as follows: *compared with control, compared with diabetic rats.
Table 7: Effect of C. auriculata seed extract on the levels of lipid profile in the plasma of experimental groups of rats
Groups
Total cholesterol
Triglycerides
LDL
HDL
Control
85.71 ± 11.54
61.89 ± 10.03
46.31 ± 5.32
28.69 ± 2.18
Diabetic
166.12 ± 18.95*
149.87 ± 16.67*
123.35 ± 8.51*
13.54 ± 1.32*
@
@
@
@
Diabetic + C.auriculata extract
107.57 ± 14.78
90.04 ± 9.85
69.72 ± 6.42
20.13 ± 1.78
@
@
@
@
Diabetic + gliclazide
96.38 ± 11.92
84.66 ± 7.59
61.07 ± 5.89
23. 51 ± 2.04
Units: mg/dl. Values are given as mean ± SD for groups of six rats in each. Values are statistically significant at p < 0.05. Statistical
@
significance was compared within the groups as follows: *compared with control, compared with diabetic rats.
The effect of oral administration of Cassia auriculata seed
extract on the levels of blood glucose, plasma insulin,
hemoglobin, glycosylated hemoglobin and urine sugar in
the control and experimental groups of rats were
depicted in Table 2. The elevated levels of blood glucose,
glycosylated hemoglobin in the diabetic group of rats
were reverted to near normal level by the administration
of Cassia auriculata seed extract. Conversely, the
decreased levels of plasma insulin, hemoglobin in diabetic
group of rats were elevated by the administration of
Cassia auriculata seed extract to diabetic rats for 30 days.
Urine sugar which is present in the diabetic group of rats
was absent in Cassia auriculata seed extract as well as
gliclazide treated diabetic group of rats. The results are
comparable with gliclazide, an oral hypoglycemic drug.
Urine sugar present in the diabetic rats was found to be
absent in the rats treated with seed extract.
The effect of oral administration of Cassia auriculata seed
extract on the levels of total protein, urea, uric acid and
creatinine are presented in Table 3. The altered levels of
these parameters were reverted back to near normalcy
upon the treatment with the seed extract.
Table 4 represents the effect of Cassia auriculata seed
extract on the levels of lipid peroxides in the plasma and
pancreas of experimental groups of rats. The levels of
lipid peroxides were significantly (p<0.05) elevated in the
diabetic group of rats. Upon oral administration of Cassia
auriculata seed extract as well as gliclazide to diabetic
group of rats were significantly (p<0.05) reverted to
normal levels when compared to control group of rats.
The effect of Cassia auriculata seed extract on the plasma
levels of non-enzymatic antioxidants such as vitamin C,
vitamin E, reduced glutathione and ceruloplasmin in
experimental groups of rats are shown in table 5. The
diminished levels of non-enzymatic antioxidants in the
diabetic group of rats were significantly (p<0.05)
improved to near normal values by the oral
administration of Cassia auriculata seed extract as well as
glyclazide, after 30 days of treatment.
Table 6 depicts the activities of pancreatic enzymatic
antioxidants such as superoxide dismutase, catalase,
glutathione peroxidase and glutathione-S-transferase in
the experimental groups of rats. The decreased activity of
enzymatic antioxidants observed in the diabetic group of
rats were significantly (p<0.05) elevated to near normal
levels after treatment with Cassia auriculata seed extract
as well as gliclazide.
The levels of cholesterol, triglycerides, HDL and LDL in
control and experimental groups of rats are shown in
Table 7. The levels of cholesterol, triglycerides, LDL and
were significantly increased whereas the HDL-cholesterol
was significantly decreased in alloxan-induced diabetic
rats. Treatment with seed extract as well as gliclazide
significantly ameliorated these levels to near normal
levels.
DISCUSSION
Alloxan, a β-cytotoxin, induces, “chemical diabetes”
through selective destruction of pancreatic beta cells
32
which results in a decrease of insulin secretion . Alloxan
also increases the oxidative stress which is the possible
33
mechanism of its diabetogenic action .
The medicinal values of plants lie in bioactive
phytochemical constituents that produce definite
physiological actions on the human body. These bioactive
phytochemical constituents in medicinal plants include
alkaloids, flavonoids, phenolic compounds, tannins,
anthracine derivatives and essential oils34. The
preliminary phytochemical screening of the Cassia
auriculata ethanolic seed extract revealed the presence
of phenols, alkaloids, flavonoids, tannins, carbohydrates.
These chemical compounds were speculated to account
for the observed pharmacological effects of the extract.
Diabetes mellitus is characterized by decreased glucose
tolerance due to low secretion of insulin or its action. This
is manifested by elevated blood glucose levels and
glucosuria, which may be accompanied by changes in lipid
and protein metabolism. When the diabetic rats were
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Volume 11, Issue 2, November – December 2011; Article-028
challenged with an oral glucose load, the blood glucose
levels reached a peak at 60 minutes and gradually
decreased to pre-glucose load level around 120 minutes,
the present study revealed that oral administration of the
ethanolic extract of Cassia auriculata seed improved the
glucose tolerance in alloxan - induced diabetic rats.
The antihyperglycemic activity of Cassia auriculata seed
extract could be due to its insulinogenic action as
increased levels of insulin were found in diabetic rats
upon treatment with extract. Therefore, the extract was
able to potentiate the release of insulin from pancreatic
islets similar to that observed after gliclazide
administration. A number of plants have been reported to
exhibit glycemic control through insulin releasing
stimulator effect35, 36.
Blood glucose is an index for the diagnosis of diabetes
mellitus. Liver functions as a “principal seat” for glucose
homeostasis and plays a vital role in the maintenance of
blood glucose levels. Alloxan administration induces
pronounced increase in the concentrations of blood
glucose. Blood glucose levels are maintained mainly by
insulin that facilitates the uptake, utilization and storage
of glucose. During diabetes, the blood glucose levels are
drastically increased which results from reduced glucose
utilization by various tissues, which is a typical condition
of insulinopenic diabetes. The elevated blood glucose
level observed in diabetic rats was almost normalized
upon extract treatment which may be due to stimulation
of glucose utilization by the peripheral tissues.
Administration of Cassia auriculata seed extract to
alloxan induced diabetic rats resulted in activation of beta
cells and granulation returns to normal insulinogenic
effect. Since the percentage fall in plasma glucose level
was different in models with varying intensity of
hyperglycemia it implies that the anti-hyperglycemic
effect of the plant is based on the dose of diabetogenic
agent and on the degree of β-cell destruction37.
Diabetes mellitus is mainly due to insulin deficiency.
Insulin deficiency is manifested in a number of
biochemical and physiological alterations. Insulin is
synthesized from its precursor proinsulin. Proinsulin is
cleaved in the secretory granules into equimolar amounts
of insulin and c-peptide38. The decreased level of the
plasma insulin observed in diabetic rats was found to be
normalized in the extract treated rats which might be due
to the insulinogenic effects of phytoconstituents present
in the seeds.
During diabetes, the excess of glucose present in blood
reacts with haemoglobin to form glycosylated
hemoglobin leads to decreased levels of hemoglobin in
39
diabetic rats . Glycation with proteins represents the
excessive levels of glucose in blood, which causes
pathophysiological changes in diabetes mellitus. In
uncontrolled diabetes, there is an increased glycation of a
number of proteins including haemoglobin and alpha
40
crystalline or lens . This glycation alters the structure and
ISSN 0976 – 044X
function of haemoglobin resulting in Hb desaturation and
precipitation of red blood cells as Heing bodies41.
Protein serves as a source of nutrition for the tissues and
its synthesis and regulation determines normal function.
Insulin plays a pivotal role in protein synthesis. Diabetes
mellitus shows profound changes in circulating
aminoacids and hepatic aminoacid uptake42. The
significant decrease of plasma protein in alloxan-induced
diabetic rats could be attributed to suppressed protein
synthesis. Urea is a non-protein nitrogenous waste
product whose level reflects a normal and continued
protein metabolism. Diabetes mellitus is associated with
changes in negative nitrogen balance and loss of nitrogen
from most organ systems. The increase in synthesis of
urea in diabetic rats may be due to the enhanced
catabolism of both liver and plasma proteins. There was
increased protein catabolism with flow of aminoacids into
43
the liver, which feeds glyconeogenesis during diabetes .
44
Dighe et al reported that accelerated proteolysis of
uncontrolled diabetes occurs as a result of deranged
glucagon mediated regulation of cAMP formation in
insulin deficiency. This accounts for the observed
decrease in the total protein content in alloxan induced
diabetic rats. Oral administration of Cassia auriculata
seed extract to diabetic rats significantly inhibits
proteolysis caused by insulin deficiency and thus
increases the level of plasma proteins to near normal
levels.
The supraphysiological concentration of glucose in
diabetic state causes severe derangement in protein
metabolism that result in the development of negative
nitrogen balance. This in turn elevates urea and
creatinine level45 (Asayama et al., 1994), which act as a
biochemical diagnostic marker for assessing renal
impairment and drug-induced toxicity. The observed
alteration in the levels of blood urea and serum creatinine
in group of diabetic rats reverted to near normalcy by
treatment with Cassia auriculata seed extract, indicating
its renal protective nature.
Uric acid is the main catabolic product from purine
nucleotides by xanthine oxidase enzymatic system. It is a
biomarker for the development of diabetic complications.
Costa et al.46 (2002) suggested increased uric acid
concentration to be a risk factor for cardiovascular
diseases. The levels of serum uric acid in diabetes induced
rats were found to be increased. Cassia auriculata extract
treatment for the diabetic rats significantly ameliorated
the toxic effects of alloxan as indicated by the restoration
of serum uric acid levels.
Diabetes is usually accompanied by increased production
of free radicals or impaired antioxidant defenses.
Excessive generation of free radicals cause damage to
cellular proteins, membrane lipids and nucleic acids and
eventually cell death. Decreased plasma insulin in diabetic
conditions increases fatty acyl coenzyme A oxidase
activity, which initiates -oxidation of fatty acids,
resulting in lipid peroxidation. Increased lipid
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peroxidation impairs membrane activity by decreasing
membrane fluidity and altering activity of membranebound enzymes and receptors. The products of lipid
peroxidation are injurious to most cells in the body and
are associated with a variety of diseases, such as
atherosclerosis and brain damage. The significant
increase in levels of TBARS observed in plasma and
pancreas of diabetic rats were decreased to near
normalcy upon oral administration of Cassia auriculata
seed extract to diabetic rats which could be a result of
improved antioxidant status.
Oxidative stress definitely refers to the situation of an
imbalance between the production of Reactive Oxygen
Species (ROS) and antioxidant defense. There is emerging
evidence that the formation of ROS is a direct
47
consequence of hyperglycemia (Daisuke koya et al.,
2003). Antioxidant treatment could be a potential
48
therapeutic procedure for diabetic complications
(Brownlee, 2001).
The enzymatic antioxidants such as SOD, catalase,
glutathione peroxidase and glutathione­S-transferase are
involved in the scavenging of reactive oxygen metabolites
that are produced as result of chronic hyperglycemia.
Gauthaman et al.49 have reported that the plant Cassia
auriculata could reduce the oxidative stress by
stimulating the activities of the scavenging enzymes like
SOD, CAT and GSH and decreasing the lipid peroxidation
levels in ischemic reperfused injuries. In diabetic milieu,
the activities of all these enzymatic antioxidants are
declined notably because of the detonated production of
free radicals arise out of chronic hyperglycemia. Oral
administration Cassia auriculata seed extract caused a
significant increase in the levels of non-enzymatic
antioxidants such as vitamin C, vitamin E, reduced
glutathione
and ceruloplasmin and enzymatic
antioxidants such as SOD, catalase, glutathione
peroxidase and glutathione-S-transferase suggesting that
the Cassia auriculata seed extract possess free radical
scavenging and antioxidant activity.
Hypertriglyceridemia and hypercholesterolemia are lipid
associated abnormalities which are characteristic features
of diabetes. Accumulation of cholesterol in liver due to
elevated plasma free fattyacids has been reported in
diabetic rats50. The higher concentration of plasma total
cholesterol observed in diabetic rats is probably due to
mobilization of free fatty acids from the peripheral fat
depots51. Cassia auriculata flower extract have reported
to possess antihyperlipidemic in triton induced
hyperlipidemic rats52.
It is well known that dyslipidemia is associated with
uncontrolled diabetes mellitus. The plasma levels of TC,
LDL-C, and TG increases, while the HDL levels decline,
53
contributing to secondary complications of diabetes .
Acute insulin deficiency initially causes and increases in
free fatty acid mobilization from adipose tissue. This
54
results in an increased production of LDL-C particle . In
the present study, diabetic rats exhibited a significant
ISSN 0976 – 044X
elevation of TC, TG and LDL-C, while HDL-C as decreased.
Cassia auriculata administration resulted in lowering the
plasma levels of TC, TG and LDL-C with elevation of HDL-C
level indicating the hypolipidemic activity of the extract.
CONCLUSION
The results of the present study indicate that the Cassia
auriculata seed extract posess both hypoglycemic,
antidyslipidemic and antioxidant activities. The possible
mechanism of the antidiabetic action may be through a
stimulation of insulin release from the remnant
pancreatic  cells. The observed antidiabetic and
antidyslipidemic effects may be due to the synergetic
actions of the phytoconstituents present in the seed
extract. The result of the present study also provides a
scientific rationale for the use of Cassia auriculata seeds
in the traditional medicine system for the treatment of
diabetes mellitus.
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